Summary Low back pain causes severe morbidity and imposes high socioeconomic burden. Degeneration of the intervertebral disc (IVD) is one of the common causes for low back pain. Cell therapy is an emerging solution to restore cellularity and functionality of the nucleus pulposus (NP) of the IVD, with potential to reverse disc degeneration or regenerate the disc. Due to poor availability, neither autologous notochordal cells nor mature NP cells are a clinically viable sources for NP regeneration. Our preliminary data showed that viable notochordal-like cells can be generated from human induced pluripotent stem cells (Liu et al. J Biomed Mater Res 2014; Liu et al. PLoS One 2014). In a stepwise manner, we coaxed iPSCs first to acquire a notochordal-like phenotype by co-expressing brachyury, cytokeratin-8 and -18 (T+/CK8+/CK18+), and then to a mature NP- like phenotype expressing SOX9, aggrecan and collagen type II, and elaborating a native-like matrix with characteristic high ratio of proteoglycan: collagen content. Based on our preliminary data, we hypothesize that human pluripotent stem cells-derived notochordal-like cells are viable progenitors for NP regeneration and yield native-like NP tissue. A central goal of this SBIR Phase I proposal is to develop a xeno-free protocol based on our existing one, and further determine the efficacy and safety of the derived notochordal-like cells in vivo. Outcome of this SBIR Phase I will establish the feasibility of a previously untapped cell source for NP regeneration, complementary to the existing work using hyaline chondrocytes and mesenchymal stromal cells.